聚合酶链反应检测与鉴定人流行性感冒病毒

Detection and Identification of Human Influenza Viruses Using Multiplex Reverse Transcrition Polumerase Chain Reaction

目的 建立一个快速、简单、敏感、特异的实验室检测流感病毒的方法。方法 采用RT-PCR、复合RT-PCR方法和HI试验对流行性感冒病毒的参考株（甲1、甲3和乙型各两株）和分离毒株（甲1型1株、甲3型3株）以及可疑流感患者的喉嗽液标本进行检测。结果 采用RT-PCR方法和复合RT-PCR方法均能检测甲1、甲3、乙型的流感病毒参考株和分离毒株,且能根据扩增物的分子量鉴定出其型别与亚型,其结果与HI法完全相符。复合RT-PCR方法从3例可疑流感患者的喉嗽液标本中检测到阳性结果1例,结果同传统鸡胚分离、HI分型方法相同。结论本文建立的RT-PCR或复合RT-PCR方法可应用于常见流感病毒的快速检测及初步分型,复合RT-PCR方法具更高的工作效率。

ve To develop a sensitive, specific, rapid method for detection and identification of human influenza viruses. Methods RT- PCR and multiplex RT-PCR were used to detect HA1, HA3, and HA(B) genes with several type or subtype specific primers in human influenza viruses. Results HA1, HA3, and HA(B) genes were able to be amplified by the RT - PCR and multiplex RT- PCR from human influenza viruses HA1, HA3, and HA(B) respectively. Virus types or subtypes were determined according to the length of amplified products, and the results were consistent with ones by hemagglutinaiton inhibition test. From 3 throat swabs samples of patients with upper respiratory tract infections, one was found HA3 gene positive by the multiplex RT-PCR, which was testified by conventional chicken embryo culture and hemagglutination inhibition test. Conclusion The RT-PCR and multiplex RT-PCR can be used in rapid detetion and identification of common human influenza viruses, and the latter is more effective.